2) OptoLoop consists of a fusion between dCas9 and the light-dependent oligomerizing protein CRY2. Genome structure can be manipulated by inducing specific chromatin loops by light. dCas9 tethers the fusion protein to target sites and CRY2:CRY2 interactions stabilize the chromatin contacts

効率的なDNA挿入技術の開発

リコンビナーゼを用いて、タンパク質進化、機械学習、dCas9との融合などによりDNA配列の効率的かつ特異的な挿入を実現。キロベース単位でのDNA挿入が可能で、幹細胞やT細胞など複数の細胞種に導入可能
www.nature.com/articles/s41...

The team developed a comprehensive engineering strategy to improve both efficiency & specificity, combining evolutionary screening to find better mutations, machine learning to predict which mutations work together & fusing the enzyme to dCas9 to guide it to the correct location.

Modeling and minimization of dCas9-induced competition in CRISPRi-based genetic circuits
doi.org/10.1101/2025...

Modeling and minimization of dCas9-induced competition in CRISPRi-based genetic circuits [new]
CRISPRi circuits limited by dCas9. Models optimize architectures mitigating these issues.

Modeling and minimization of dCas9-induced competition in CRISPRi-based genetic circuits https://www.biorxiv.org/content/10.1101/2025.11.05.686856v1

Modeling and minimization of dCas9-induced competition in CRISPRi-based genetic circuits https://www.biorxiv.org/content/10.1101/2025.11.05.686856v1

By coupling the CRISPR/dCas9 based SunTag system with the plant H3K4me3 methyltransferase, SDG2, we show that H3K4me3 can activate the expression of a gene that is typically silent, FWA. Works super well, and honestly slightly unexpected as DNA methylation is though to be the main player for FWA...

We also show combinatorial genetic and epigenetic engineering with CRISPR Cas12a-dCas9 to perform targeted CAR knock-in with CRISPRoff-silencing of target genes to improve preclinical CAR T performance. Read the full paper here: rdcu.be/eL0GK

A preprint using: pHR-SFFV-dCas9-BFP (RRID:Addgene_46910) from @addgene.bsky.social was published.

SciScore made a table with this resource, see “Automated Services” module (download as csv, xml or #jats) #RRID #OpenScience

A preprint using: pHR-SFFV-dCas9-BFP (RRID:Addgene_46910) from @addgene.bsky.social was published.

SciScore made a table with this resource, see “Automated Services” module (download as csv, xml or #jats) #reproducibility #RRID

I suggested:
- look for dCas9 by IF and Western blot
- qPCR something from the plasmid
Am I missing something?

I can imagine! Congrats again to you and the whole team on this beautiful piece of work. Can I ask a technical question please: why did you choose a cumate-inducible dCas9-KRAB instead of for example Dox inducible?

3️⃣ Future synthetic biology utilities:

Right now, we are fusing these proteins to #synbio effectors and adaptors to:
• control RNA fate,
• boost CRISPR/dCas9 RNA scaffolds,
• reduce off-target labeling
enabling next-gen RNA–protein circuits for synthetic regulation and control.

CrAAVe-seq delivers sgRNA libraries via AAV vectors into LSL-dCas9-KRAB mice. In these mice, only Cre+ neurons excise the Stop cassette and express dCas9-KRAB. The sgRNAs then guide dCas9-KRAB to DNA, silencing the target genes.

dCas9 targeted proteome profiling reveals p300-mediated reciprocal regulation of SMAD and SP1 as a driver of GM2 synthase transcription in renal cell carcinoma https://www.biorxiv.org/content/10.1101/2025.09.16.676212v1

dCas9 targeted proteome profiling reveals p300-mediated reciprocal regulation of SMAD and SP1 as a driver of GM2 synthase transcription in renal cell carcinoma https://www.biorxiv.org/content/10.1101/2025.09.16.676212v1

Real-Time Imaging of Specific Genomic Loci With CRISPR/dCas9 in Human Cells Using CRISPRainbow

Work by Thomas J. Versosky, Dilshodbek U. Nishonov and Li-Chun Tu at Ohio State University.

#LifeSciences #Reproducibility #Biophysics